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目的观察PTD-HBcAg融合蛋白免疫BALB/c小鼠后的体液免疫与细胞免疫效果。方法目的蛋白(PTD-HBcAg)、阳性对照蛋白(HBcAg)和阴性对照分别与等体积的弗氏完全佐剂乳化免疫小鼠,第21天以不完全弗氏佐剂乳化同法加强免疫,第28天ELISA法检测HBcAb滴度,流式细胞仪检测HBV特异性IFN-γ+、CD8+T淋巴细胞水平。结果PTD-HBcAg融合蛋白能够诱导产生较高水平HBcAb;与阳性蛋白对照组[(1.45±0.19)%,(2.19±0.04)%和(4.32±1.04)%]和阴性对照组相比,PTD-HBcAg融合蛋白组[(2.64±0.21)%,(3.50±0.50)%和(6.85±1.3)%]诱导的HBV特异性CD8+T淋巴细胞水平明显增高(P<0.05)。结论PTD-HBcAg融合蛋白具有体液免疫与细胞免疫效果,为治疗性乙肝疫苗的开发提供了一定的实验基础。
Objective To observe the humoral and cellular immune responses of BALB / c mice immunized with PTD-HBcAg fusion protein. Methods The target protein (PTD-HBcAg), positive control protein (HBcAg) and negative control were respectively immunized with an equal volume of Freund’s complete adjuvant. The mice were immunized on the 21st day with incomplete Freund’s adjuvant emulsification. HBcAb titers were detected by ELISA for 28 days, and the levels of HBV-specific IFN-γ + and CD8 + T lymphocytes were detected by flow cytometry. Results PTD-HBcAg fusion protein could induce a higher level of HBcAb. Compared with the positive control group [(1.45 ± 0.19)%, (2.19 ± 0.04)% and (4.32 ± 1.04)%] The level of HBV specific CD8 + T lymphocytes induced by HBcAg fusion protein was significantly higher than that of HBcAg fusion protein [(2.64 ± 0.21)%, (3.50 ± 0.50)% and (6.85 ± 1.3)%], respectively. Conclusion PTD-HBcAg fusion protein has humoral and cellular immunity effects, which provides a certain experimental basis for the development of therapeutic hepatitis B vaccine.